Thermal activation of single kinesin molecules with temperature pulse microscopy

Kenji Kawaguchi, ShiN'Ichi Ishiwata

    Research output: Contribution to journalArticle

    40 Citations (Scopus)

    Abstract

    Conventional kinesin is a processive motor protein that keeps "walking" along a microtubule using chemical energy released by ATP hydrolysis. We previously studied the effects of temperature between 15° and 35°C on the moving velocity, force, and processivity of single kinesin molecules using a bead assay [Kawaguchi and Ishiwata, 2000b: Biochem Biophys Res Commun 272:895-899]. However, we could not examine the effects of temperature higher than 35°C because of the thermal damage to proteins. Here, using temperature pulse microscopy (TPM) [Karo et al., 1999: Proc Natl Acad Sci USA 96:9602-9606], we could examine the temperature dependence of the gliding velocity of single kinesin molecules interacting with a microtubule above 35°C up to 50°C (instantaneously, ∼60°C), where the velocity reached 3.68 μm/s, the highest ever reported. The Arrhenius plot showed no breaks between 15° and 50°C with a unique activation energy of about 50 kJ/mol, suggesting that the molecular mechanism of kinesin motility is common over a broad temperature range including physiological temperature.

    Original languageEnglish
    Pages (from-to)41-47
    Number of pages7
    JournalCell Motility and the Cytoskeleton
    Volume49
    Issue number1
    DOIs
    Publication statusPublished - 2001

    Fingerprint

    Kinesin
    Microscopy
    Hot Temperature
    Temperature
    Microtubules
    Walking
    Proteins
    Hydrolysis
    Adenosine Triphosphate

    Keywords

    • Arrhenius plot
    • Microtubule
    • Motor protein
    • Single molecule assay
    • Temperature effect on kinesin

    ASJC Scopus subject areas

    • Cell Biology

    Cite this

    Thermal activation of single kinesin molecules with temperature pulse microscopy. / Kawaguchi, Kenji; Ishiwata, ShiN'Ichi.

    In: Cell Motility and the Cytoskeleton, Vol. 49, No. 1, 2001, p. 41-47.

    Research output: Contribution to journalArticle

    Kawaguchi, Kenji ; Ishiwata, ShiN'Ichi. / Thermal activation of single kinesin molecules with temperature pulse microscopy. In: Cell Motility and the Cytoskeleton. 2001 ; Vol. 49, No. 1. pp. 41-47.
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